Anti-polyubiquitin antibodies and methods of use

Abstract

The invention provides anti-polyubiquitin antibodies and methods of using the same.

What is claimed is: 
     
       1. An isolated monoclonal antibody that binds K11-linked polyubiquitin, wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 2 and 57 to 60, an HVR-L2 sequence selected from SEQ ID NOs: 3 and 61, an HVR-L3 sequence of SEQ ID NO: 4, an HVR-H1 sequence selected from SEQ ID NOs: 6 to 11, an HVR-H2 sequence selected from SEQ ID NOs: 12 to 17 and 67, and an HVR-H3 sequence selected from SEQ ID NOs: 18 to 23, 68 and 69. 
     
     
       2. The antibody of  claim 1 , wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 2 and 57 to 60, an HVR-L2 sequence selected from SEQ ID NOs: 3 and 61, an HVR-L3 sequence of SEQ ID NO: 4, an HVR-H1 sequence of SEQ ID NO: 11, an HVR-H2 sequence selected from SEQ ID NOs: 17 and 67, and an HVR-H3 sequence selected from SEQ ID NOs: 23, 68 and 69. 
     
     
       3. The antibody of  claim 1  , wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 58 and 59, an HVR-L2 sequence of SEQ ID NO: 3, an HVR-L3 sequence of SEQ ID NO: 4, an HVR-H1 sequence of SEQ ID NO: 11, an HVR-H2 sequence of SEQ ID NO: 67, and an HVR-H3 sequence of SEQ ID NO: 23. 
     
     
       4. The antibody of  claim 1 , wherein the antibody comprises an HVR-L1 sequence of SEQ ID NO: 59, an HVR-L2 sequence of SEQ ID NO:3, an HVR-L3 sequence of SEQ ID NO: 4, an HVR-H1 sequence of SEQ ID NO: 11, an HVR-H2 sequence of SEQ ID NO: 67, and an HVR-H3 sequence of SEQ ID NO: 23. 
     
     
       5. The antibody of  claim 1 , wherein the antibody comprises a heavy chain variable region comprising a sequence with at least 95% sequence identity to SEQ ID NO. 72 and a light chain variable region comprising a sequence with at least 95% sequence identity to SEQ ID NO: 65. 
     
     
       6. The antibody of  claim 5 , wherein the antibody comprises a heavy chain variable region comprising the sequence of SEQ ID NO. 72 and a light chain variable region of SEQ ID NO: 65. 
     
     
       7. (The antibody of  claim 1 , comprising light chain and heavy chain amino acid sequences with at least 95% sequence identity to the amino acid sequences of one of the following combinations of sequences: SEQ ID NOs: 5 and 32; SEQ ID NOs: 63 and 32; SEQ ID NOs: 65 and 32; SEQ ID NOs: 5 and 72; SEQ ID NOs: 63 and 72; and SEQ ID NOs: 65 and 72; SEQ ID NOs: 62 and 32; SEQ ID NOs: 64 and 32; SEQ ID NOs: 5 and 70; SEQ ID NOs: 5 and 71; SEQ ID NOs: 66 and 32. 
     
     
       8. The antibody of  claim 7 , comprising light chain and heavy chain amino acid sequences of one of the following combinations of sequences: SEQ ID NOs: 5 and 32; SEQ ID NOs: 63 and 32; SEQ ID NOs: 65 and 32; SEQ ID NOs: 5 and 72; SEQ ID NOs: 63 and 72; and SEQ ID NOs: 65 and 72; SEQ ID NOs: 62 and 32; SEQ ID NOs: 64 and 32; SEQ ID NOs: 5 and 70; SEQ ID NOs: 5 and 71; SEQ ID NOs: 66 and 32. 
     
     
       9. The isolated antibody of  claim 1 , wherein the antibody specifically binds to a K11-linked polyubiquitinated protein. 
     
     
       10. The isolated antibody of  claim 1 , wherein the antibody modulates at least one polyubiquitin-mediated signaling pathway. 
     
     
       11. The isolated antibody of  claim 1 , wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody, or an antibody fragment that binds K11-linked polyubiquitin. 
     
     
       12. The antibody of  claim 1 , wherein the antibody binds K11-linked polyubiquitin under stringent conditions, but does not bind K48-linked polyubiquitin under stringent conditions, wherein the stringent conditions comprise 4M urea. 
     
     
       13. The antibody of  claim 1 , wherein the antibody binds K11-linked polyubiquitin under stringent conditions, but does not bind K63-linked polyubiquitin under stringent conditions, wherein the stringent conditions comprise 4M urea. 
     
     
       14. The antibody of  claim 1 , wherein the antibody binds K11-linked polyubiquitin under stringent conditions, but does not bind K48-linked polyubiquitin or K63-linked polyubiquitin under stringent conditions, wherein the stringent conditions comprise  4 M urea. 
     
     
       15. The antibody of  claim 1 , wherein the antibody does not bind monoubiquitin. 
     
     
       16. An immunoconjugate comprising the antibody of  claim 1  and a cytotoxic agent. 
     
     
       17. A pharmaceutical formulation comprising the antibody of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
       18. The pharmaceutical formulation of  claim 17 , further comprising an additional therapeutic agent. 
     
     
       19. An isolated monoclonal antibody that binds to the same antigenic determinant on K11-linked ppolyubiquitin as the antibody of  claim 6 . 
     
     
       20. An isolated monocianal antibody that competes with the antibody of  claim 6  for binding to K11-linked polyubiquitin.